LOCUS       sequence-252712-        5654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express hNABP1 N-terminus (1–141 aa) in E. coli with a C-terminal 
            His-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5654)
  AUTHORS   Vidhyasagar V, He Y, Guo M, Ding H, Talwar T, Nguyen V, Nwosu J, 
            Katselis G, Wu Y
  TITLE     C-termini are essential and distinct for nucleic acid binding of 
            human NABP1 and NABP2.
  JOURNAL   Biochim Biophys Acta. 2016 Feb;1860(2):371-83. doi: 
            10.1016/j.bbagen.2015.11.003. Epub 2015 Nov 10.
  PUBMED    26550690
REFERENCE   2  (bases 1 to 5654)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.bbagen.2015.11.003"; journalName: "Biochim Biophys Acta";
            date: "2016-02"; volume: "1860"; issue: "2"; pages: "371-83"
COMMENT     Sequence Label: pET29a-hNABP1-N
FEATURES             Location/Qualifiers
     source          1..5654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     488..507
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(527..604)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     810..829
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     917..936
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        917..935
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    936..960
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             975..997
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1431..1448
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1501..1519)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1515..1562
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1599..2054
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1686..1705)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1896..1917
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2146..2961)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2146..2961)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2869..2888
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2953..2972
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3083..3671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3572..3591
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3825..3842
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3857..3996
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3922..3941)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4082..4104
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4098..4289)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5098..5654,1..526))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
       61 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
      121 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
      181 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
      241 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
      301 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
      361 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
      421 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
      481 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
      541 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
      601 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
      661 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
      721 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
      781 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
      841 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat
      901 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
      961 cctctagaaa taattttgtt taactttaag aaggagatat acatatgaat agggtcaacg
     1021 acccacttat ttttataaga gatattaagc ccggactgaa aaacttaaat gtcgtcttta
     1081 ttgtcctgga gataggacgc gtgaccaaaa ccaaagacgg ccatgaagtg agatcgtgca
     1141 aagtagcaga taaaacgggc agcatcacta tttccgtgtg ggatgagatc ggaggtctta
     1201 tacagccagg ggatattatt cggttgacca gagggtatgc atccatgtgg aaaggatgtc
     1261 tgacacttta tactggaagg ggtggtgaac ttcaaaaaat tggggaattt tgtatggttt
     1321 attcagaagt gccaaatttc agtgaaccca acccagatta tcgaggacag cagaacaaag
     1381 gggcacagag tgaacagaag aataattcca tgaatagtaa tatgctcgag caccaccacc
     1441 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     1501 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1561 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
     1621 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     1681 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     1741 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     1801 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     1861 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     1921 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     1981 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2041 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2101 tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc
     2161 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     2221 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     2281 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     2341 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     2401 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     2461 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     2521 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     2581 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     2641 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     2701 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     2761 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     2821 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     2881 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt
     2941 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     3001 tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3061 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3121 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3181 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3241 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3301 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3361 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3421 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3481 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3541 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3601 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3661 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3721 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3781 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3841 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3901 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     3961 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     4021 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4081 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     4141 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     4201 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     4261 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     4321 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     4381 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     4441 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     4501 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     4561 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     4621 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     4681 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     4741 atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     4801 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     4861 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     4921 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     4981 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     5041 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     5101 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5161 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     5221 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     5281 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     5341 tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa cgcgcagccc
     5401 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     5461 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     5521 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     5581 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     5641 ctggtgaccc aatg
//
